本文選題：雷公藤紅素 + 多西紫杉醇��； 參考：《哈爾濱工業(yè)大學(xué)》2016年碩士論文

【摘要】：前列腺癌(Prostate Cancer,PCa)是在男性中發(fā)病率和死亡率都非常高的癌癥之一。雖然隨著檢測手段的更新以及治療方式的多樣化,但是大部分病人一旦發(fā)展出轉(zhuǎn)移瘤,進而轉(zhuǎn)變?yōu)镃RPC(Castration-resistant PCa),最終產(chǎn)生藥物抗性時,其生存期將大大降低。CRPC被認為與AR(Androgen Receptor)通路的再激活關(guān)系密切,AR通路的再激活是導(dǎo)致腫瘤存活的關(guān)鍵。AR剪切變異體AR-V7不需要雄激素激活轉(zhuǎn)運入核,能夠激活A(yù)R通路調(diào)控下游基因。因此,AR-V7已成為治療CRPC前列腺癌的腫瘤治療的研究熱點。多西紫杉醇作為治療CRPC的一線化療藥物,能夠通過抑制微管解聚進而抑制AR核轉(zhuǎn)入并激活細胞凋亡。然而CRPC最終會發(fā)展出藥物抗性,抗性來源可能是凋亡通路的改變以及AR-V7的表達。雷公藤紅素作為一種天然的蛋白酶體抑制劑,能夠通過誘導(dǎo)細胞凋亡、抑制AR的表達進而抑制前列腺癌細胞系的生長。因此,本研究利用AR-V7陽性細胞系22Rv1作為模型,并將AR-V7陰性細胞系LNCa P作為對照,分析并解釋多西紫杉醇與雷公藤紅素對前列腺癌細胞的生長抑制作用差異以及其機理。本研究通過MTT實驗確證了22Rv1為多西紫杉醇抗性細胞系、LNCa P則為敏感細胞系,同時與雷公藤紅素聯(lián)合用藥后細胞存活率相似,聯(lián)合用藥計算顯示雷公藤紅素能夠恢復(fù)22Rv1對多西紫杉醇的敏感性,而LNCa P經(jīng)過藥物聯(lián)用處理后則表現(xiàn)出拮抗效果。凋亡實驗顯示,0.5 nmol/L的多西紫杉醇與0.8μmol/L的雷公藤紅素單獨用藥對AR-V7表達差異的兩株前列腺癌細胞誘導(dǎo)的凋亡結(jié)果無明顯差異,藥物聯(lián)用下22Rv1的凋亡率上升至50.62%,LNCa P則僅為32.6%。這說明了雷公藤紅素在AR-V7陽性細胞系中能恢復(fù)多西紫杉醇誘導(dǎo)凋亡的能力。同時Western Blot結(jié)果顯示,雷公藤紅素能夠恢復(fù)多西紫杉醇在多西紫杉醇抗性細胞系中的PARP切割,并且在多西紫杉醇處理下22Rv1細胞系較LNCa P細胞PARP切割表達量少,這說明了雷公藤紅素能夠通過恢復(fù)多西紫杉醇抗性細胞系內(nèi)已改變的凋亡通路產(chǎn)生增敏作用。q RT-PCR結(jié)果顯示,雷公藤紅素能夠從m RNA水平上抑制AR、AR-V7的表達,而多西紫杉醇不能夠影響其表達。這說明,雷公藤紅素能夠通過抑制AR、AR-V7表達抑制細胞內(nèi)的AR通路。綜上,本研究初步分析雷公藤紅素與多西紫杉醇對前列腺癌細胞生長抑制差異,闡明雷公藤紅素抑制AR-V7陽性細胞生長、藥物增敏的部分分子機制。
[Abstract]:Prostate Cancer (PCa) is one of the cancers with high morbidity and mortality among men. Although with the renewal of detection methods and the diversification of the methods of treatment, the survival period will be large when most patients develop metastatic tumors and then turn to CRPC (Castration-resistant PCa) and eventually produce drug resistance. .CRPC is believed to be closely related to the reactivation of the AR (Androgen Receptor) pathway, and the reactivation of the AR pathway is the key.AR shear variant AR-V7 that does not require androgen activation to enter the nucleus, and can activate the AR pathway to regulate the downstream genes. Therefore, AR-V7 has become a research heat for the treatment of cancer of CRPC prostate cancer. As a first-line chemotherapeutic agent for the treatment of CRPC, docetaxel can inhibit microtubule disaggregation and inhibit the transfer of AR nuclei to activate cell apoptosis. However, CRPC eventually develops drug resistance, and the source of resistance may be a change in the apoptosis pathway and the expression of AR-V7. In this study, the AR-V7 positive cell line 22Rv1 was used as a model, and the AR-V7 negative cell line LNCa P was used as the control, and the difference in the inhibitory effect of docetaxel and tripterin on the growth of the adenocarcinoma cells of the anterior adenocarcinoma cells was analyzed and explained, as well as the inhibition of the growth of the prostate cancer cell lines by inducing apoptosis. Therefore, the AR positive cell line 22Rv1 was used as a model. Mechanism. In this study, the MTT test confirmed that 22Rv1 was a docetaxel resistant cell line, LNCa P was a sensitive cell line, and the cell survival rate was similar to that of tripterin. The combined use of tripterin could restore the sensitivity of 22Rv1 to docetaxel, while LNCa P was treated with a combination of drugs. The apoptosis test showed that 0.5 nmol/L of docetaxel and 0.8 mol/L of tripterin alone had no significant difference in the apoptosis induced by two prostate cancer cells with different AR-V7 expression, the apoptosis rate of 22Rv1 increased to 50.62%, and LNCa P was only 32.6%., which indicated that tripterin in AR-V7 In the positive cell line, the ability to induce docetaxel to induce apoptosis was restored. Western Blot results showed that tripterine could restore the PARP cutting of docetaxel in the docetaxel resistant cell line, and the PARP cut expression in 22Rv1 cell lines was less than that of LNCa P cells under docetaxel treatment, which indicated that tripterine was reproduced. .q RT-PCR can be produced by restoring the altered apoptotic pathway in the cell line of the docetaxel resistant cell line. The results show that Lei Gongteng erythropoietin can inhibit the expression of AR and AR-V7 from m RNA level, while docetaxel can not affect its expression. This shows that Lei Gongteng erythropoietin can inhibit AR by inhibiting AR and AR-V7 expression. To sum up, this study preliminarily analyzed the difference between tripterine and docetaxel on the growth inhibition of prostate cancer cells, and elucidated the molecular mechanism of tripterine inhibiting the growth of AR-V7 positive cells and sensitizing the drugs.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R737.25
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本文編號：1993973